CN102965415B - Regioselective fucosylation modification method of enzymatic nuclear glucoside class medicine - Google Patents

Regioselective fucosylation modification method of enzymatic nuclear glucoside class medicine Download PDF

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CN102965415B
CN102965415B CN201210470287.5A CN201210470287A CN102965415B CN 102965415 B CN102965415 B CN 102965415B CN 201210470287 A CN201210470287 A CN 201210470287A CN 102965415 B CN102965415 B CN 102965415B
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beta
damping fluid
fucoside
nucleosides
galactosidase enzymes
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CN102965415A (en
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李宁
宗敏华
颜丽强
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South China University of Technology SCUT
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Abstract

The invention discloses a regioselective fucosylation modification method of an enzymatic nuclear glucoside class medicine, and the method comprises the following specific steps of: (1) evenly mixing a buffer solution, nucleoside and p-nitro-phenyl-beta-fucoside together; (2) adding beta-galactosidase in an enzyme dosage of 1-30 U/gram of nucleoside, reacting for 5-100 hours under the conditions of 30-55 DEG C and 200 r/min, treating for 10 minutes at 100 DEG C, and performing quenching enzyme reaction; and (3) obtaining the modified product through column chromatography isolation. The method has the advantages of simple reaction process, high regioselectivity, no complex protection and deprotection steps, moderate reaction conditions, environmental friendliness, easiness for separation of products and the like.

Description

A kind of method that enzyme catalysis nucleoside medicine regioselectivity fucosylation is modified
Technical field
The invention belongs to applying biological catalytic field, be specifically related to a kind of method of utilizing beta-galactosidase enzymes catalysis nucleoside medicine regioselectivity fucosylation to modify.
Background technology
Nucleoside medicine is the important antitumor and Anti-virus agent of a class (J Clin Virol, 2004,30,115).For example, FdUrd has been widely used in the tumours such as treatment colorectal carcinoma and hepatic metastases, and acyclovir is used to treat herpesvirus infection.Yet nucleoside medicine shows lower oral administration biaavailability conventionally, therefore need heavy dose of and often take to keep Plasma Concentration higher than the required lowest threshold for the treatment of.In addition, this class drug selectivity is poor, when killing sick cell, also normal cell is had to stronger toxicity, therefore conventionally show larger toxic side effect (Br J Pharmacol, 2006,147,1) in clinical use.
Structure activity study shows; through glycosylation modified synthetic glycosides derivatives, can effectively shield the toxicity of female medicine; when this analog derivative arrives after pathological tissues; hydrolytic action by beta-glycosidase in pathological tissues born of the same parents, discharges female medicine, thereby effectively improves female medicine selectivity; reduce its toxic side effect (Bioorg Med ChemLett; 2007,17,983; Angew Chem Int Ed, 2009,48,6443).For example, Abraham etc. find that 5-FUD is after glycosyl galactose is modified, and the secondary work of the poison of Balb/c mouse has been reduced to nearly 100 times (CellBiochem Biophys, 1994,24,127).It is toxicity (J Med Chem, 1981,24,893) that 5 '-glucuronide derivative of the discovery 5-FUDs such as Watanabe and 5-fluorine cytidine also can greatly shield female medicine.Meanwhile, glycosides derivatives, by the identification with small intestine inwall glucokinase activity, semi-lactosi carrier proteins, can effectively strengthen the intestinal absorption of medicine by active transport, improves its bioavailability (J Pharm Sci, 1998,87,326).And, the glycosylation modified stability that can also effectively improve medicine, water-soluble etc.For example, the stability of two kinds of glycosides derivatives of the discovery such as Bonina zidovudine in blood, far above female medicine, has significantly improved the retention time of zidovudine in blood, thereby greatly reduced administration number of times (Eur J Pharm Sci, 2002,16,167).
In addition, the fucoside derivative of nucleosides is the important building block of many bioactive natural products.For example inosine fucoside derivative is the important synthon that preparation has the Shimofuridin A of stronger resisting gram-positive bacteria and anti-mycotic activity; Guanosine fucoside derivative is the synthetic front extract (Russ Chem Rev, 2004,73,401) with the HF-7 of stimulating neural tissue activity.
At present, two ribosides mainly synthesize by chemical method, comprise two kinds of approach: a) O-glycosylation: by nucleosides and monose, through O-glycosylation, obtained; B) N-glycosylation: obtain (Mol Biol, 2009,43,301) through N-glycosylation by disaccharides and nucleoside base.Owing to all having a plurality of activity hydroxies or amino in two substrates, the poor selectivity of chemical method, thus synthetic protection and the deprotection steps that needs multistep much time power of two ribosides, complex process.Meanwhile, when purine biosynthesis class two riboside, need to use highly toxic mercury salt catalyzer, environment unfriendly (Russ Chem Rev, 2004,73,401).
The synthetic biocatalysis approach of two ribosides is still very limited, and only has report (J Mol Catal B:Enzym, 2007,47,28 of enzymatic nucleosides galactosylation and glucosyl reaction; J Mol Catal B:Enzym, 2012,79,35; Bioorg Med Chem, 2012,20,3111; J Biotechnol, 2011,155,203).Up to the present, still fail to realize synthetic two ribosides containing β-fucosido of enzyme catalysis nucleosides regioselectivity fucosylation reaction.
Summary of the invention
The special substrate specificity that beta-galactosidase enzymes based on deriving from beef liver, bull testis and pork liver has, the problem existing for prior art, the invention provides a kind of method of utilizing beta-galactosidase enzymes catalysis nucleoside medicine regioselectivity fucosylation to modify synthetic nucleosides fucoside derivative.
Technical solution of the present invention is as follows:
(1) damping fluid, nucleosides and p-nitrophenyl β-fucoside are mixed;
(2) ratio that is 1 ~ 30U/ gram of nucleosides in enzyme dosage adds beta-galactosidase enzymes, under 30 ~ 55 ℃, 200r/min, reacts after 5 ~ 100h, processes 10min, cancellation enzyme reaction at 100 ℃;
(3) after centrifugal, utilize liquid chromatogram measuring production concentration, according to product calibration curve, calculate productive rate, or except after desolventizing, the product after utilizing column chromatography for separation to be modified.
Preferably, the described nucleosides of step (1) and p-nitrophenyl β-fucoside mol ratio are 5:1 ~ 1:4.
Preferably, the described beta-galactosidase enzymes of step (2) is the beta-galactosidase enzymes that derives from beef liver, bull testis and pork liver.
Preferably, the pH value of the described damping fluid of step (1) is 5.0 ~ 9.0.
Preferably, the described damping fluid of step (1) is citric acid-Na 2hPO 4damping fluid or glycine-NaOH damping fluid.
Preferably, the described nucleosides of step (1) is a kind of in 2 '-deoxyuridine, FdUrd, the chloro-2 '-deoxyuridine of 5-, the bromo-2 '-deoxyuridine of 5-, iodoxuridine, beta-thymidine, Trifluridine, 5-FUD, zidovudine, stavudine, inosine, guanosine, acyclovir, ganciclovir and Penciclovir.Structural formula is as follows:
Figure BDA00002425925800031
R＝H，F，Cl，Br，I，CH 3，CF 3
Figure BDA00002425925800032
Y＝H，NH 2 X＝CH 2，O
The present invention compared with prior art has advantages of as follows:
1) the present invention is in damping fluid, take p-nitrophenyl β-fucoside as glycosyl donor, utilizes beta-galactosidase enzymes catalysis nucleoside medicine to carry out regioselectivity glycosylation, synthetic nucleosides fucoside derivative.This enzyme reaction has height regioselectivity, therefore overcome traditional chemical method selectivity low, easily generate by product, need protection and deprotection operation, the shortcoming such as productive rate is low and environment is unfriendly.
2) the present invention is without radical protection and deprotection operation, and synthesis technique is simple, and product is easily separated.
3) the present invention is under temperature is 30 ~ 55 ℃, condition of normal pressure, utilizes enzymatic modification to obtain fucoside derivative, and reaction conditions is gentle, environmental friendliness.
Accompanying drawing explanation
Fig. 1 is the NMR hydrogen spectrogram of 5 '-O-β-fucosido-FdUrd.
Fig. 2 is the NMR carbon spectrogram of 5 '-O-β-fucosido-FdUrd.
Fig. 3 is the NMR hydrogen spectrogram of 5 '-O-β-fucosido-2 '-deoxyuridine.
Fig. 4 is the NMR carbon spectrogram of 5 '-O-β-fucosido-2 '-deoxyuridine.
Fig. 5 is the NMR hydrogen spectrogram of the bromo-2 '-deoxyuridine of 5 '-O-β-fucosido-5-.
Fig. 6 is the NMR carbon spectrogram of the bromo-2 '-deoxyuridine of 5 '-O-β-fucosido-5-.
Fig. 7 is the NMR hydrogen spectrogram of 5 '-O-β-fucosido-iodoxuridine.
Fig. 8 is the NMR carbon spectrogram of 5 '-O-β-fucosido-iodoxuridine.
Fig. 9 is the NMR hydrogen spectrogram of 5 '-O-β-fucosido-acyclovir.
Figure 10 is the NMR carbon spectrogram of 5 '-O-β-fucosido-acyclovir.
Embodiment
By embodiment, further illustrate the present invention, but be not limited to embodiment.
Embodiment 1
In 15mL triangular flask with cover, add 2mL citric acid-Na 2hPO 4damping fluid (50mM, pH 7.0), 0.02mmol p-nitrophenyl β-fucoside and 0.04mmol FdUrd, mix, and adds subsequently 0.06U beef liver beta-galactosidase enzymes, under 40 ℃ and 200r/min, reacts.After 90h, reaction solution is warming up to 100 ℃ and processes 10min, cancellation enzyme reaction, the centrifugal rear concentration of utilizing liquid chromatogram measuring product 5 '-O-β-fucosido-FdUrd, according to product calibration curve, calculating productive rate is 30%.Product NMR hydrogen spectrum and carbon spectrum are respectively as shown in attached Fig. 1 and 2.
Embodiment 2
In 15mL triangular flask with cover, add 2mL citric acid-Na 2hPO 4damping fluid (50mM, pH 6.0), 0.02mmol p-nitrophenyl β-fucoside and 0.04mmol FdUrd, mix, and adds subsequently 0.21U beef liver beta-galactosidase enzymes, under 40 ℃ and 200r/min, reacts.After 24h, reaction solution is warming up to 100 ℃ and processes 10min, cancellation enzyme reaction, the centrifugal rear concentration of utilizing liquid chromatogram measuring product 5 '-O-β-fucosido-FdUrd, according to product calibration curve, calculating productive rate is 41%.
Embodiment 3
In 15mL triangular flask with cover, add 2mL citric acid-Na 2hPO 4damping fluid (50mM, pH 7.0), 0.02mmol p-nitrophenyl β-fucoside and 0.04mmol FdUrd, mix, and adds subsequently 0.21U beef liver beta-galactosidase enzymes, under 50 ℃ and 200r/min, reacts.After 17h, reaction solution is warming up to 100 ℃ and processes 10min, cancellation enzyme reaction, the centrifugal rear concentration of utilizing liquid chromatogram measuring product 5 '-O-β-fucosido-FdUrd, according to product calibration curve, calculating productive rate is 40%.
Embodiment 4
In 15mL triangular flask with cover, add 2mL citric acid-Na 2hPO 4damping fluid (50mM, pH 7.0), 0.02mmol p-nitrophenyl β-fucoside and 0.10mmol FdUrd, mix, and adds subsequently 0.21U beef liver beta-galactosidase enzymes, under 40 ℃ and 200r/min, reacts.After 27h, reaction solution is warming up to 100 ℃ and processes 10min, cancellation enzyme reaction, the centrifugal rear concentration of utilizing liquid chromatogram measuring product 5 '-O-β-fucosido-FdUrd, according to product calibration curve, calculating productive rate is 53%.
Embodiment 5
In 15mL triangular flask with cover, add 2mL citric acid-Na 2hPO 4damping fluid (50mM, pH 7.0), 0.02mmol p-nitrophenyl β-fucoside and 0.10mmol FdUrd, mix, and adds subsequently 0.30U bull testis beta-galactosidase enzymes, under 45 ℃ and 200r/min, reacts.After 20h, reaction solution is warming up to 100 ℃ and processes 10min, cancellation enzyme reaction, the centrifugal rear concentration of utilizing liquid chromatogram measuring product 5 '-O-β-fucosido-FdUrd, according to product calibration curve, calculating productive rate is 50%.
Embodiment 6
In 15mL triangular flask with cover, add 2mL citric acid-Na 2hPO 4damping fluid (50mM, pH 7.0), 0.02mmol p-nitrophenyl β-fucoside and 0.06mmol 2 '-deoxyuridine, mix, and adds subsequently 0.42U pork liver beta-galactosidase enzymes, under 40 ℃ and 200r/min, reacts.After 24h, reaction solution is warming up to 100 ℃ and processes 10min, cancellation enzyme reaction, the centrifugal rear concentration of utilizing liquid chromatogram measuring product 5 '-O-β-fucosido-2 '-deoxyuridine, according to product calibration curve, calculating productive rate is 43%.Product NMR hydrogen spectrum and carbon spectrum are respectively as shown in accompanying drawing 3 and 4.。
Embodiment 7
In 15mL triangular flask with cover, add 2mL citric acid-Na 2hPO 4damping fluid (50mM, pH 7.0), 0.02mmol p-nitrophenyl β-fucoside and the bromo-2 '-deoxyuridine of 0.06mmol 5-, mix, and adds subsequently 0.21U beef liver beta-galactosidase enzymes, under 40 ℃ and 200r/min, reacts.After 24h, reaction solution is warming up to 100 ℃ and processes 10min, cancellation enzyme reaction, the centrifugal rear concentration of utilizing the bromo-2 '-deoxyuridine of liquid chromatogram measuring product 5 '-O-β-fucosido-5-, according to product calibration curve, calculating productive rate is 44%.Product NMR hydrogen spectrum and carbon spectrum are respectively as shown in accompanying drawing 5 and 6.。
Embodiment 8
In 15mL triangular flask with cover, add 2mL citric acid-Na 2hPO 4damping fluid (50mM, pH 7.0), 0.02mmol p-nitrophenyl β-fucoside and 0.06mmol iodoxuridine, mix, and adds subsequently 0.21U beef liver beta-galactosidase enzymes, under 40 ℃ and 200r/min, reacts.After 24h, reaction solution is warming up to 100 ℃ and processes 10min, cancellation enzyme reaction, the centrifugal rear concentration of utilizing liquid chromatogram measuring product 5 '-O-β-fucosido-iodoxuridine, according to product calibration curve, calculating productive rate is 60%.Product NMR hydrogen spectrum and carbon spectrum are respectively as shown in accompanying drawing 7 and 8.
Embodiment 9
In 15mL triangular flask with cover, add 2mL citric acid-Na 2hPO 4damping fluid (50mM, pH 7.0), 0.02mmol p-nitrophenyl β-fucoside and 0.06mmol acyclovir, mix, and adds subsequently 0.21U beef liver beta-galactosidase enzymes, under 40 ℃ and 200r/min, reacts.After 48h, reaction solution is warming up to 100 ℃ and processes 10min, cancellation enzyme reaction, the centrifugal rear concentration of utilizing liquid chromatogram measuring product 4 '-O-β-fucosido-acyclovir, according to product calibration curve, calculating productive rate is 36%.Product NMR hydrogen spectrum and carbon spectrum are respectively as shown in accompanying drawing 9 and 10.
Embodiment 10
In 250mL triangular flask with cover, add 40mL citric acid-Na 2hPO 4damping fluid (50mM, pH7.0), 0.4mmol p-nitrophenyl β-fucoside and 0.8mmol FdUrd, mix, and adds subsequently 2.1U beef liver beta-galactosidase enzymes, under 40 ℃ and 200r/min, reacts.After 24h, reaction solution is warming up to 100 ℃ and processes 10min, cancellation enzyme reaction, except after desolventizing, utilizes column chromatography for separation to obtain 71mg 5 '-O-β-fucosido-FdUrd (45% yield).
Embodiment 11
In 250mL triangular flask with cover, add 40mL citric acid-Na 2hPO 4damping fluid (50mM, pH7.0), 0.4mmol p-nitrophenyl β-fucoside and 1.2mmol iodoxuridine, mix, and adds subsequently 2.1U beef liver beta-galactosidase enzymes, under 40 ℃ and 200r/min, reacts.After 24h, reaction solution is warming up to 100 ℃ and processes 10min, cancellation enzyme reaction, except after desolventizing, utilizes column chromatography for separation to obtain 118mg 5 '-O-β-fucosido-iodoxuridine (59% yield).
Embodiment 12
In 250mL triangular flask with cover, add 40mL citric acid-Na 2hPO 4damping fluid (50mM, pH7.0), 0.4mmol p-nitrophenyl β-fucoside and 1.2mmol acyclovir, mix, and adds subsequently 2.1U beef liver beta-galactosidase enzymes, under 40 ℃ and 200r/min, reacts.After 48h, reaction solution is warming up to 100 ℃ and processes 10min, cancellation enzyme reaction, except after desolventizing, utilizes column chromatography for separation to obtain 49mg 5 '-O-β-fucosido-acyclovir (33% yield).

Claims (6)

1. the method that enzyme catalysis nucleoside medicine regioselectivity fucosylation is modified, is characterized in that, concrete steps are as follows:
(1) damping fluid, nucleosides and p-nitrophenyl β-fucoside are mixed;
(2) ratio that is 1 ~ 30U/ gram of nucleosides in enzyme dosage adds beta-galactosidase enzymes, under 30 ~ 55 ℃, 200r/min, reacts after 5 ~ 100h, processes 10min, cancellation enzyme reaction at 100 ℃;
(3) product after utilizing column chromatography for separation to be modified.
2. method according to claim 1, it is characterized in that, the described nucleosides of step (1) is a kind of in 2 '-deoxyuridine, FdUrd, the chloro-2 '-deoxyuridine of 5-, the bromo-2 '-deoxyuridine of 5-, iodoxuridine, beta-thymidine, Trifluridine, 5-FUD, zidovudine, stavudine, inosine, guanosine, acyclovir, ganciclovir and Penciclovir.
3. method according to claim 1 and 2, is characterized in that, the described nucleosides of step (1) and p-nitrophenyl β-fucoside mol ratio are 5:1 ~ 1:4.
4. method according to claim 1 and 2, is characterized in that, the pH value of the described damping fluid of step (1) is 5.0 ~ 9.0.
5. method according to claim 4, is characterized in that, described damping fluid is citric acid-Na 2hPO 4damping fluid or glycine-NaOH damping fluid.
6. method according to claim 1 and 2, is characterized in that, the described beta-galactosidase enzymes of step (2) is the beta-galactosidase enzymes that derives from beef liver, bull testis and pork liver.
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EP3082456A1 (en) * 2013-12-11 2016-10-26 DuPont Nutrition Biosciences ApS A method for preparing a dairy product having a stable content of galacto-oligosaccharide(s)
CN105315318B (en) * 2015-11-06 2019-04-19 山东大学 A kind of alpha-L-Rhamnosidase is preparing the application in the fluoro- 2 '-deoxidation urea glycoside derivates of 5-
CN105418711A (en) * 2015-11-06 2016-03-23 山东大学 Application of alpha-L-rhamnosidase to preparing hydroxycarbamide and glycoside derivatives
CN107236010A (en) * 2017-06-28 2017-10-10 山东大学 A kind of application of α L rhamnosidases in cytarabine derivative is prepared

Citations (2)

* Cited by examiner, † Cited by third party
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CN101778937A (en) * 2007-06-04 2010-07-14 诺和诺德公司 o-linked glycosylation using n-acetylglucosaminyl transferases
CN102037004A (en) * 2008-01-08 2011-04-27 生物种属学股份公司 Glycoconjugation of polypeptides using oligosaccharyltransferases

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101778937A (en) * 2007-06-04 2010-07-14 诺和诺德公司 o-linked glycosylation using n-acetylglucosaminyl transferases
CN102037004A (en) * 2008-01-08 2011-04-27 生物种属学股份公司 Glycoconjugation of polypeptides using oligosaccharyltransferases

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
Highly regioselective glucosylation of 2’-deoxynucleosides by using the crude β-glycosidase from bovine liver;Min Ye et al.;《Journal of Biotechnology》;20110910;第155卷(第2期);第204页左栏第1段和材料与方法部分第2.5小节 *
Min Ye et al..Highly regioselective glucosylation of 2’-deoxynucleosides by using the crude β-glycosidase from bovine liver.《Journal of Biotechnology》.2011,第155卷(第2期),第203–208页.

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